Anna Arvidsson

Nano hydroxyapatite structures influence early bone formation

In a study model that aims to evaluate the effect of nanotopography on bone formation, micrometer structures known to alter bone formation, should be removed. Electropolished titanium implants were prepared to obtain a surface topography in the absence of micro structures, thereafter the implants were divided in two groups. The test group was modified with nanosize hydroxyapatite particles; the other group was left uncoated and served as control for the experiment. Topographical evaluation demonstrated increased nanoroughness parameters for the nano-HA implant and higher surface porosity compared to the control implant. The detected features had increased size and diameter equivalent to the nano-HA crystals present in the solution and the relative frequency of the feature size and diameter was very similar. Furthermore, feature density per microm(2) showed a decrease of 13.5% on the nano-HA implant. Chemical characterization revealed calcium and phosphorous ions on the modified implants, whereas the control implants consisted of pure titanium oxide. Histological evaluation demonstrated significantly increased bone formation to the coated (p < 0.05) compared to uncoated implants after 4 weeks of healing. These findings indicate for the first time that early bone formation is dependent on the nanosize hydroxyapatite features, but we are unaware if we see an isolated effect of the chemistry or of the nanotopography or a combination of both.

Bone reaction to nano hydroxyapatite modified titanium implants placed in a gap-healing model

Nanohydroxyapatite materials show similar chemistry to the bone apatite and depending on the underlying topography and the method of preparation, the nanohydroxyapatite may simulate the specific arrangement of the crystals in bone. Hydroxyapatite (HA) and other CaP materials have been indicated in cases in which the optimal surgical fit is not achievable during surgery, and the HA surface properties may enhance bone filling of the defect area. In this study, very smooth electropolished titanium implants were used as substrata for nano-HA surface modification and as control. One of each implant (control and nano HA) was placed in the rabbit tibia in a surgical site 0.7 mm wider than the implant diameter, resulting in a gap of 0.35 mm on each implant side. Implant stability was ensured by a fixating plate fastened with two side screws. Topographical evaluation performed with an optical interferometer revealed the absence of microstructures on both implants and higher resolution evaluation with AFM showed similar nanoroughness parameters. Surface pores detected on the AFM measurements had similar diameter, depth, and surface porosity (%). Histological evaluation demonstrated similar bone formation for the nano HA and electropolished implants after 4 weeks of healing. These results do not support that nano-HA chemistry and nanotopography will enhance bone formation when placed in a gap-healing model. The very smooth surface may have prevented optimal activity of the material and future studies may evaluate the synergic effects of the surface chemistry, micro, and nanotopography, establishing the optimal parameters for each of them.

Chemical and topographical analyses of dentine surfaces after Carisolv™ treatment

OBJECTIVES The aim of this study was to characterise the surface chemistry of cavities after chemomechanical caries excavation, and also to measure the surface topography after caries removal with Carisolv or burs, followed by acid etching. METHODS Fourier transform (FT)-Raman spectroscopy was used to study the relative amounts of organic material and minerals of sound enamel, dentine, and cavities, after caries excavation. Fourier transform infrared spectroscopy (FTIR) and laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) were used for detection of Carisolv substances (i.e. mainly sodium hypochlorite, amino acids, and the gelling agent carboxymethyl cellulose). In total, 19 carious and 11 sound extracted teeth were used for the chemical analyses. Topographic examination of 30 carious extracted teeth was performed with a contact profilometer. RESULTS The relative amounts of organic material and minerals did not significantly differ between sound dentine and the cavities after caries removal with burs or Carisolv. The FTIR analyses indicated extremely small amounts of Carisolv substances at the cavity surface, but the LA-ICP-MS analyses did not confirm those findings. Furthermore, the topographical parameters did not significantly differ between etched cavities after caries removal using burs or Carisolv. CONCLUSIONS The chemical and topographical analyses in the present study imply that any differences between the cavities after caries excavation with burs or with Carisolv are insignificant.

An in vitro comparison of possibly bioactive titanium implant surfaces

The aim of the study was to compare Ca and P formation (CaP) and subsequent bone cell response of a blasted and four different possibly bioactive commercially pure (cp) titanium surfaces; 1. Fluoride etched (Fluoride), 2. Alkali-heat treated (AH), 3. Magnesium ion incorporated anodized (TiMgO), and 4. Nano HA coated and heat treated (nano HA) in vitro. Furthermore, to evaluate the significance of the SBF formed CaP coat on bone cell response. The surfaces were characterized by Optical Interferometry, Scanning Electron Microscopy (SEM) and X-ray Photoelectron Spectroscopy (XPS). CaP formation was evaluated after 12, 24 and 72 h in simulated body fluid (SBF). Primary human mandibular osteoblast-like cells were cultured on the various surfaces subjected to SBF for 72 h. Cellular attachment, differentiation (osteocalcin) and protein production (TGF-beta(1)) was evaluated after 3 h and 10 days respectively. Despite different morphological appearances, the roughness of the differently modified surfaces was similar. The possibly bioactive surfaces gave rise to an earlier CaP formation than the blasted surface, however, after 72 h the blasted surface demonstrated increased CaP formation compared to the possibly bioactive surfaces. Subsequent bone cell attachment was correlated to neither surface roughness nor the amount of formed CaP after SBF treatment. In contrast, osteocalcin and TGF-beta(1) production were largely correlated to the amount of CaP formed on the surfaces. However, bone response (cell attachment, osteocalcin and TGF-F production) on the blasted controls were similar or increased compared to the SBF treated fluoridated, AH and TiMgO surface.

Early interactions between leukocytes and three different potentially bioactive titanium surface modifications

The aim of the present study was to compare the early interactions between leukocytes and three different surface modifications, suggested as bioactive. Blasted titanium discs were modified by alkali and heat treatment, sodium fluoride treatment, or hydroxyapatite coating. A number of these discs were also immersed in simulated body fluid (SBF) for a week, a treatment which yielded high levels of calcium and phosphate on each surface type. The specimens were exposed for human venous blood for 32 minutes and the respiratory burst response was measured in terms of reactive oxygen species with a luminometer, and coverage of viable cells with a fluorescence microscope after staining steps. The topography, morphology, and chemistry of the surfaces were evaluated with optical interferometry and scanning electron microscopy/energy dispersive X-ray analysis (SEM/EDX). A high respiratory burst response was found for HA coated titanium in comparison with the other surface groups (p < 0.0005). The SBF immersion resulted in an increased respiratory burst response (p < 0.0005) and removed statistically significant differences between the surface groups. Thus, the results in the present study indicate that different titanium surface modifications influence the early inflammatory response differently, and that calcium phosphate compounds increase the inflammatory response.

Comparing and visualizing titanium implant integration in rat bone using 2D and 3D techniques

The aim was to compare the osseointegration of grit-blasted implants with and without a hydrogen fluoride treatment in rat tibia and femur, and to visualize bone formation using state-of-the-art 3D visualization techniques. Grit-blasted implants were inserted in femur and tibia of 10 Sprague-Dawley rats (4 implants/rat). Four weeks after insertion, bone implant samples were retrieved. Selected samples were imaged in 3D using Synchrotron Radiation-based μCT (SRμCT). The 3D data was quantified and visualized using two novel visualization techniques, thread fly-through and 2D unfolding. All samples were processed to cut and ground sections and 2D histomorphometrical comparisons of bone implant contact (BIC), bone area (BA), and mirror image area (MI) were performed. BA values were statistically significantly higher for test implants than controls (p < 0.05), but BIC and MI data did not differ significantly. Thus, the results partly indicate improved bone formation at blasted and hydrogen fluoride treated implants, compared to blasted implants. The 3D analysis was a valuable complement to 2D analysis, facilitating improved visualization. However, further studies are required to evaluate aspects of 3D quantitative techniques, with relation to light microscopy that traditionally is used for osseointegration studies.

Characterisation of Structures in Salivary Secretion Film Formation. An Experimental Study with Atomic Force Microscopy

The purpose of the present study was to characterise the structure dynamics of pure salivary secretions retained on controlled surfaces with different surface energies in the early stage of salivary film formation. Germanium prisms prepared to have either low surface energy or medium surface energy were incubated in fresh secretions of either human parotid saliva (HPS) or human submandibular/sublingual saliva (HSMSLS) for 15, 90, and 180 min. After controlled rinsing with distilled water, the surfaces were air dried and thereafter imaged with atomic force microscopy (AFM). The amount of adsorbed material and the size of the structures detected increased with increased saliva exposure time. The film thicknesses varied from 10 to 150 nm, and both HPS and HSMSLS films contained structures with diameters varying from 40 nm to 2 μm. Some of these were clustered into special formations. The HPS films exhibited a more granular morphology than the HSMSLS films. Furthermore, branched lines were detected on the low surface energy germanium prisms incubated in saliva. The results indicate that exposure time, surface energy, and type of salivary secretion all are factors affecting the adsorption characteristics of salivary films.

Influence of chemo‐mechanical caries removal on the surface topography of dental composite resin and glass‐ionomer materials: an in vitro study

The aim of the present study was to investigate the influence of a chemo‐mechanical caries removal system, Carisolv ® gel, on the surface topography of dental filling materials. Thirty specimens of a composite resin (Spectrum™) and a compomer (Dyract ® AP) and 60 specimens of a glass‐ionomer (Ketac ® ‐Fil Plus) were prepared. The surface topography was investigated with an optical interferometer before and after chemical exposure for 5, 10, or 20 min. Each specimen acted as its own control. The topographical part of the glass‐ionomer materials was performed in two series with different drying procedures, since this material exhibits a higher sensitivity to dehydration than the other materials. The surface topographical investigations were complemented with contact angle measurements. After Carisolv ® gel exposure the density of summits and the developed surface area ratio (3D/2D) were slightly smaller for the composite resin and the compomer. For the minimally dried glass‐ionomer material, the results indicated a reduction of the height of the surface structures, as well as a surface area enlargement, after Carisolv ® gel exposure. No statistically significant changes of contact angles due to Carisolv ® gel exposure could be detected for any material investigated. If dental filling materials of composite resin or glass‐ionomer materials are exposed to Carisolv ® gel, no or only minor surface topographical changes can be expected.